pCS2+hSpCas9
(Plasmid #51815)

• Purpose: Encode hSpCas9 for in vitro transcription with Sp6
• Depositing Lab: Masato Kinoshita
• Publication: Ansai et al Biol Open. 2014 Apr 11;3(5):362-71. doi: 10.1242/bio.20148177.

Backbone

• Vector backbone: pCS2
• Backbone size w/o insert (bp): 4071
• Total vector size (bp): 8343
• Modifications to backbone: Cloned in hSpCas9 expression sequence
• Vector type: Mammalian Expression, CRISPR ; medaka, zebrafish

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: humanized S.pyogenes Cas9
• Alt name: SpCas9
• Alt name: hSpCas9
• Insert Size (bp): 4272
• Promoter: SP6
• Tag / Fusion Protein:
  • 3xFLAG (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: Xba I (not destroyed)
• 5′ sequencing primer: SP6
• 3′ sequencing primer: T3

Resource Information

• Supplemental Documents:
  • pCS2+hSpCas9 genebank annotated file
• A portion of this plasmid was derived from a plasmid made by: Dr. Zhang produced the hSpCas9 gene, which has been deposited to Addgene (Plasmid 42230: pX330-U6-Chimeric_BB-CBh-hSpCas9 | Cong et al. Science. 3 Jan 2013).
• Articles Citing this Plasmid:
  • 29 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid is modified from pX330-U6-Chimeric BB-CBh-hSpCas9 originated from Feng Zhang Lab.